Properties of the catalase molecule obtained from acatalasemic and hypocatalasemic mice Part II. Analysis of liver catalase of acatalasemic mice by isoelectrofocusing.

The properties of catalases in the liver homogenate of acatalasemic (C3H/C(sb)C(sb)), hypocatalasemic (C3H/C(sc)C(sc)), acatalasemic heterozygous (C3H/C(sa)C(sb)) and normal (C3H/C(sa)C(sa)) mice were analyzed directly by agarose isoelectric focusing.Homogenates of mouse liver were prepared with isotonic sucrose solution and separated from the subcellular fractions by repeated centrifugation followed by ultra-centrifugation (105,000×g).Agarose isoelectric focusing was performed in a pH 3-to-10 Pharmalyte gradient gel at 8℃.Isoelectric range of catalase in liver supernatants was compared normal and mutant mice which were designated acatalasemia, hypocatalasemia and acatalasemic heterozygous.Liver catalases of C(sb)C(sb),C(sc)C(sc), C(sa)C(sb) and C(sa)C(sa) were focused to a band at pl 5.9-7.6,pl 5.8-7.3, pl 5.6-7.0 and pl 5.4-7.0, respectively. The difference in the isoelectric points of catalases among C(sb)C(sb), C(sc)C(sc), C(sa)C(sb) and C(sa)C(sb) suggested the presence of some structural differences in the catalase molecule between normal and mutant mice.The homegenates of liver from C(sa)C(sa) and C(sb)C(sb) were separated into four fractions (non-catalase fraction and A,B and C catalase fractions) by DEAE cellulose column chromatography with a discontinous buffer system of 1mM, 3mM, 10mM and 100mM phosphate buffer. The eluates were concentrated to 1.5 PU/ml by ultrafiltration, and pl values of the fractions were examined by isoelectric focusing. Acatalasemic fractions B and C focused at a more alkaline pl than normal fractions B and C.